Wetzel D M, Bohn M C, Hamill R W
Department of Neurology, University of Rochester Medical Center, Monroe Community Hospital, NY 14620.
Brain Res. 1994 Jun 27;649(1-2):117-21. doi: 10.1016/0006-8993(94)91054-5.
The relative postmortem stability of the mRNA's for glucocorticoid (GR) and mineralocorticoid (MR) receptor in rodent brain was determined using semi-quantitative in situ hybridization (ISH). Rats were killed by CO2 asphyxiation and their brains removed immediately (0 h) or following 12 h or 24 h delays. Specific hybridization of GR and MR anti-sense [35S]RNA-probe to tissue mRNA encoding these receptors was detected using film and emulsion autoradiography. The most intense labeling for GR mRNA was in the dentate gyrus followed by the CA1 hippocampal region. Lower, but still detectable signal, was apparent over CA3-CA4 pyramidal cell regions. MR mRNA was detected throughout the CA1-4 pyramidal cell fields of the hippocampus and the granular cells of the dentate gyrus. Film images demonstrated that even in the 24 h postmortem delay group intense specific signal was present in sections hybridized with both anti-sense GR and MR probes, although there was some diminution in signal intensity in cortical areas at this later postmortem delay. These initial experiments with rat brain demonstrate that the mRNA's for both GR and MR, as detected with ISH, are stable for up to 24 h following death.
使用半定量原位杂交(ISH)技术测定了啮齿动物脑中糖皮质激素(GR)和盐皮质激素(MR)受体mRNA的相对死后稳定性。大鼠通过二氧化碳窒息处死,其大脑在立即(0小时)或延迟12小时或24小时后取出。使用胶片和乳胶放射自显影检测GR和MR反义[35S]RNA探针与编码这些受体的组织mRNA的特异性杂交。GR mRNA的最强标记出现在齿状回,其次是海马CAI区。在CA3 - CA4锥体细胞区域也有较低但仍可检测到的信号。在整个海马的CA1 - 4锥体细胞区域和齿状回的颗粒细胞中检测到MR mRNA。胶片图像显示,即使在死后延迟24小时的组中,与反义GR和MR探针杂交的切片中仍存在强烈的特异性信号,尽管在这个较晚的死后延迟时皮质区域的信号强度有所减弱。这些对大鼠脑的初步实验表明,用ISH检测到的GR和MR的mRNA在死后长达24小时内是稳定的。